Expression and induction of cytochrome P-450 1A1 and P-450 2D subfamily in the rat glioma C6 cell line.
The cytochrome P-450 (P-450) monooxygenase system can catalyze the oxidation of a wide variety of endogenous and exogenous compounds, including steroid hormones, fatty acids, drugs and pollutants. The functions of this system are as diverse as the substrates. Though this enzyme system has the highest level of activity in the liver, it is present in other tissues, including brain. In this study, we have established the rat glioma C6 cell line as an in vitro model system to examine the expression and induction of P-450 1A1 and the P-450 2D subfamily. Rat glioma C6 cells were treated with P-450 inducers phenobarbital (PB) or benzo[a]anthracene (BA). The presence of P-450 1A1 and 2D1-5 was detected by reverse transcription followed by polymerase chain reaction (RT-PCR) and confirmed by restriction enzyme digestion. The induction of P-450 1A1 and 2D1-5 was quantified using competitive PCR. Although P-450 2D1-5 do not seem to be affected by PB or BA treatment, tenfold induction of P-450 1A1 mRNA after BA treatment was detected. Western blot analysis of microsomal preparations of glioma C6 cells demonstrated the presence of P-450 1A1 at the protein level. ELISAs showed that BA induces P-450 1A1 proteins 7.3-fold. These experiments provide further evidence that the rat glioma C6 cell line contains an active cytochrome P-450 monooxygenase system which can be induced by P-450 inducers. In summary, we believe the presence of the cytochrome P-450 monooxygenase system in glial cells of the brain may be important in chemotherapy and carcinogenesis of brain tumors.
Geng
J
J
Department of Biochemistry and Molecular Biology, The University of Texas Medical School, Houston 77225, USA.
Strobel
H W
HW
D000818:Animals; D001551:Benz(a)Anthracenes / Q000494:pharmacology; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D005910:Glioma / Q000378:metabolism* / Q000473:pathology; D008297:Male; D010634:Phenobarbital / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014407:Tumor Cells, Cultured
D001551:Benz(a)Anthracenes; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; C030935:benz(a)anthracene; D019363:Cytochrome P-450 CYP1A1; D010634:Phenobarbital